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A novel calcium-dependent chloride current in Xenopus oocytes injected with brain messenger RNA.

1. Membrane currents were studied in voltage-clamped Xenopus laevis oocytes which had been injected with total rat brain RNA. 2. When the membrane potential was stepped from -100 to +10 mV, two components of outward current were observed which were named Tout1 and Tout2. 3. Both Tout1 and Tout2 were eliminated in chloride-free or calcium-free media and blocked by 9-anthroic acid, indicating that they represented calcium-dependent chloride currents. 4. Both currents were dependent on extracellular calcium (1.8-10 mM), with Tout1 showing a greater sensitivity to changes in calcium concentration. 5. Tout2 but not Tout1 was blocked by intracellular injection of 300-600 pmol, BaCl2 (final concentration in the oocyte: 0.3-0.6 mM). Injection of KCl had no effect on either Tout1 or Tout2. 6. Tout2 but not Tout1 was enhanced by low concentrations of serotonin (0.5-2 nM). This effect was blocked by 0.1 microM-mianserin. Higher concentrations (above 10 nM) of serotonin decreased the amplitude of Tout2. The effect of serotonin was blocked by the protein kinase inhibitor, H-7 (25 microM). 7. Tout2 but not Tout1 was enhanced by 10 nM-phorbol myristate acetate. Higher concentrations of the phorbol ester decreased the amplitude of Tout2. 8. It is concluded that in oocytes injected with RNA there is an induction of a novel component of the calcium-induced chloride current (Tout2). This current reflects a second process of chloride channel opening which can be enhanced by serotonin via activation of protein kinase C.

Animals↗

The effect of a standard oral calcium load given in two different forms on plasma ionized calcium and serum PTH.

A standard dose (1 g, 25.4 mmol) of calcium was administered both as calcium chloride in syrup and as Calcium-Sandoz Syrup (calcium glubionate and calcium galactogluconate) to 10 volunteers. Both dosage forms caused a significant rise in ionized calcium and a significant fall in concentration of serum PTH, the calcium chloride producing significantly greater changes. The urinary excretion of calcium measured over 5 h after calcium chloride was double that after Calcium-Sandoz. Calcium chloride, as expected, also gave rise to a decreased urine pH. Calcium chloride would be the better choice for an oral PTH suppression test.

Administration, Oral↗

Consumer acceptance of calcium chloride-marinated top loin steaks.

In this study, a multi-city, retail consumer survey was used to determine whether marinating in calcium chloride at 72 h postmortem improves consumer and trained sensory panel evaluations of beef loin steaks, Warner-Bratzler shear force (WBSF) values, and the value of CaCl2 marinades to improve tenderness in a national consumer market. Boneless strip loin subprimals (n = 8) were obtained from eight, not electrically stimulated, USDA Standard-grade carcasses, halved, and alternating ends were marinated with 200 mM CaCl2 at 5% (wt/wt) or not injected (controls). Strip loins were then aged for 7 d in vacuum packages at 2 degrees C before WBSF, trained or consumer evaluations. Four trained research teams traveled on the same 7-d period to collect the consumer data. Consumers (n = 393; minimum of 10 consumers/panel, three panels/store, three stores/city, and four cities) varying widely in income level, education, and gender evaluated the steaks for tenderness, beef flavor, juiciness, and overall quality in each of three supermarkets in major cities (Los Angeles, CA; Baltimore, MD; Chicago, IL; and Dallas, TX). The average improvement (P < 0.05) in WBSF for steaks marinated with CaCl2 was 1.39 kg. Trained panelists scored marinated steaks higher (P < 0.05) than control steaks in sustained juiciness (6.00 vs. 5.09), beef flavor (6.31 vs. 5.64), and overall mouth-feel (5.98 vs. 4.93). Consumers also rated marinated steaks higher (P < 0.05) than control steaks in juiciness (5.98 vs. 5.45), beef flavor (6.70 vs. 6.05), tenderness (6.75 vs. 5.89), and overall quality (6.68 vs. 6.20). Consumers in this study would pay dollar 0.95/kg more for steaks marinated with CaCl2, adding dollar 21.64 to the value of a loin from tough carcasses. These results indicate that consumers can differentiate tenderness levels and are willing to pay a premium for calcium chloride marinating to effectively improve the quality of beef loin steaks.

Adult↗

Hypercalcemia attenuates blood pressure response to epinephrine.

The potent alpha and beta adrenergic actions of epinephrine are probably mediated through an increase in free intracellular calcium concentration. Despite an absence of experimental evidence, many clinicians administer epinephrine with calcium to augment its cardiovascular effects. We evaluated the effects of calcium on epinephrine's pressor response in both normal and endotoxin-treated rats by administering epinephrine to animals made hypercalcemic with calcium chloride or hypocalcemic with the calcium chelator EGTA. EPI, given in incremental doses of 10, 20 and 50 micrograms/kg, produced incremental increases in mean arterial pressure. Calcium chloride infused i.v. at a rate of 50 mg/ml/hr significantly (P less than .05) blunted (50% decrease) the hypertensive response to 50 micrograms/kg epinephrine in normal animals. In endotoxin-treated rats, calcium chloride at 50 mg/ml/hr significantly blunted the hypertensive response to 10 micrograms/kg (73% decrease), 20 micrograms/kg (62% decrease) and 50 micrograms/kg (50% decrease) epinephrine. Endotoxemia plus calcium chloride at 25 mg/ml/hr also significantly blunted (30% decrease) the hypertensive response to 50 micrograms/kg EPI. By contrast, hypocalcemia produced by EGTA (30 mg/ml/hr) had no effect on epinephrine's hypertensive effects in normal or endotoxemic rats. Since calcium chloride significantly diminishes epinephrine's hypertensive effects in both normal and endotoxin-treated rats, the clinical use of calcium chloride along with epinephrine may not have a sound experimental basis.

Animals↗

Calcium influx requirement for human neutrophil chemotaxis: inhibition by lanthanum chloride.

Calcium fluxes of human neutrophils measured in the presence of chemotactically active serum showed a marked stimulation of calcium-45 uptake from the media. Chemotactically inactive serum did not cause an influx of calcium. The magnitude of the calcium influx due to activated serum is sufficient to trigger contractile systems previously described in muscle cells. Lanthanum chloride inhibited the chemotactic response of human neutrophils to activated serum. Lanthanum in concentrations that suppressed chemotaxis also inhibited the calcium influx caused by activated serum. Lanthanum in concentrations that suppressed chemotaxis also inhibited the calcium influx caused by activated serum. The data support a direct role of calcium influx in chemotaxis of neutrophils.

Biological Transport↗

Expression of hepatic calcium-binding protein regucalcin mRNA is mediated through Ca2+/calmodulin in rat liver.

The effect of signal transduction-related factors was investigated to clarify the expression mechanism for mRNA of the hepatic Ca(2+)-binding protein regucalcin in the liver of rats. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin cDNA (0.6 kb). A single intraperitoneal administration of calcium chloride (15 mg Ca2+: 0.374 mmol/100 g body weight) to rats induced a remarkable increase of regucalcin mRNA in liver; the level was about 170% of controls at 30 min after administration. This increase was completely inhibited by simultaneous administration of trifluoperazine (5.0 mg/100 g), an antagonist of calmodulin. On the other hand, a single intraperitoneal administration of phorbol ester or dibutyryl cAMP (10-1,000 micrograms/100 g) did not cause a significant alteration of hepatic regucalcin mRNA levels. Also, administration of zinc, copper and cadmium (0.374 mmol of metal ion/100 g) did not have an appreciable effect on hepatic regucalcin mRNA levels. These findings demonstrate that the expression of hepatic regucalcin mRNA is mediated through Ca2+/calmodulin.

Animals↗

Extracellular zinc stimulates a calcium-activated chloride conductance through mobilisation of intracellular calcium in renal inner medullary collecting duct cells.

We have used the perforated patch clamp and fura-2 fluorescence techniques to study the effect of extracellular Zn(2+) on whole-cell Ca(2+)-activated Cl(-) currents (I (CLCA)) in mouse inner medullary collecting duct cells (mIMCD-3). I (CLCA) was spontaneously active in 74% of cells under basal conditions and displayed time and voltage-independent kinetics and an outwardly rectifying current/voltage relationship (I/V). Addition of zinc chloride (10-400 microM) to the bathing solution resulted in a dose-dependent increase in I (CLCA) with little change in Cl(-) selectivity or biophysical characteristics, whereas gadolinium chloride (30 microM) and lanthanum chloride (100 microM) had no significant effect on the whole-cell current. Using fura-2-loaded mIMCD-3 cells, extracellular Zn(2+) (400 microM) stimulated an increase in intracellular Ca(2+) to an elevated plateau. The Zn(2+)-stimulated [Ca(2+)](i) increase was inhibited by thapsigargin (200 nM), the IP(3) receptor antagonist 2-aminoethoxydiphenyl borate (10 microM) and removal of bath Ca(2+). Pre-exposure to Zn(2+) (400 microM) markedly attenuated the ATP (100 microM)-stimulated [Ca(2+)](i) increase. These data are consistent with the hypothesis that extracellular Zn(2+) stimulates an increase in [Ca(2+)](i) by a release of calcium from thapsigargin/IP(3) sensitive stores. A possible physiological role for a divalent metal ion receptor, distinct from the extracellular Ca(2+)-sensing receptor, in IMCD cells is discussed.

Animals↗

EFFECT OF SALINITY AND TEMPERATURE ON COCCIDIOIDES IMMITIS AND THREE ANTAGONISTIC SOIL SAPROPHYTES.

Egeberg, Roger O. (University of Southern California, Los Angeles), Ann F. Elconin, and Margaret C. Egeberg. Effect of salinity and temperature on Coccidioides immitis and three antagonistic soil saprophytes. J. Bacteriol. 88:473-476. 1964.-Experiments exploring the factors which determine the ability of Coccidioides immitis to survive and reproduce in the soil in situ are described and discussed. Previous work indicated that soil temperatures and salinity influence the growth and distribution of C. immitis. It was thought that these factors might work through enhancing the growth of C. immitis or inhibiting the growth of its possible antagonists, or both. In searching for antagonists, samples of surface soil from an endemic area were screened by standard culture techniques. Three significant antagonists were isolated and were identified as two strains of Bacillus subtilis, and Penicillium janthinellum. Subcultures of each of these isolates were grown on Saboraud's media modified by sodium chloride and calcium chloride in parallel series. The salts used were the predominant salts, and the range of temperature was comparable to that found in the soil. The organisms were grown at 18 to 27 C and at 40 C. The latter temperature approximated the temperature of the top centimeter of soil for 4 to 6 hr a day in the spring when C. immitis is recovered. P. janthinellum was unable to survive 40 C. The growth of the other two, while unaffected by temperature change alone, was inhibited by the salts. Conversely, C. immitis was stimulated by the salts when grown at the higher temperature. These findings support the hypothesis that markedly increased salinity and seasonal high temperature of the surface soil inhibit or kill these antogonists while enhancing the growth of C. immitis.

Bacillus subtilis↗

Normalization of priming solution ionized calcium concentration improves hemodynamic stability of neonates receiving venovenous ECMO.

The authors' objectives in this investigation were: 1) to prospectively determine whether a normocalcemic priming solution would result in elimination of hypocalcemia after the initiation of extracorporeal membrane oxygenation (ECMO); 2) to investigate whether normocalcemia would result in improvements in the patient's hemodynamics during the initiation of ECMO; and 3) to further define the relationship between ionized calcium measurements and total calcium, serum total protein, serum albumin, and total magnesium. This was a prospective study done in our neonatal intensive care unit, and included nine neonatal patients placed on ECMO for cardiopulmonary support. The bypass circuit was primed in the standardized manner with 100 mg calcium chloride. Circuit ionized calcium measurements were performed, and additional calcium chloride was added to normalize the ionized calcium in the priming solution. Ionized calcium was measured from the circuit and the patient before the initiation of ECMO, and then again from the patient 5, 10, 15, 30, 60, 120, and 240 minutes after the initiation of ECMO. The patients' mean arterial pressure was measured simultaneously with each ionized calcium measurement. Ionized calcium, serum total calcium, total protein, serum albumin, and total magnesium were measured from blood samples simultaneously collected four times daily. There was no significant change in the ionized calcium measured in the patients after the initiation of ECMO. There was, however, a significant increase in blood pressure 5 min after the initiation of ECMO (62 +/- 7 mmHg vs 53 +/- 6 mmHg, p = 0.01). Thereafter, there was no difference in blood pressure measured when compared with pre ECMO values. A poor correlation was demonstrated between ionized calcium and total calcium (r2 = 0.35), serum total protein (r2 = 0.26), serum albumin (r2 = 0.27), and total magnesium (r2 = 0.05). On the basis of the authors' data, the initiation of ECMO with a normocalcemic prime results in a minimal change in patient ionized calcium and resolution of the hypotension previously observed. In addition, there was poor correction between ionized calcium, total calcium, and other indirect measures of ionized calcium. Ionized calcium measurements are critical for patient hemodynamic stability before bypass and should be normalized in both the patient and priming solution before the initiation of bypass.

Blood Pressure↗

Calcium-activated chloride channels: (un)known, (un)loved?

Calcium-activated chloride channels (CaCCs) participate in many different physiologic processes such as transepithelial transport, excitability of neurons and muscle cells, and oocyte fertilization. Within the airways, they contribute to epithelial fluid secretion. This review focuses on three outstanding questions about CaCCs. First, although their biophysical fingerprint (anion selectivity, Ca2+ and voltage dependence, kinetics) is fairly well established, the molecular identity of CaCCs is still unresolved. CLCA, a family of proteins of which four members have so far been identified in humans, has been proposed to mediate calcium-activated chloride currents. However, the biophysical profile and expression pattern of endogenous CaCCs differ from those of the CLCA proteins. Another family of membrane proteins, the bestrophins, has recently been shown in transfected HEK293 cells to confer anion-selective currents that are activated by submicromolar Ca2+ concentrations. Second, pharmacologic tools to manipulate CaCCs are poorly selective. This lack of specificity not only hampers the structural and functional characterization of these channels but also restricts therapeutic options for altering CaCC function. Finally, potential pitfalls with respect to CaCCs as molecular targets for cystic fibrosis therapy are discussed.

Animals↗

Inhibition of growth of human breast cancer cell line MCF-7 by serum derived from calcium chloride-clotted plasma.

The growth of MCF-7 cells, established from a metastatic mammary carcinoma, and of HBL-100 cells, derived from a primary culture of human milk, was examined in medium supplemented with whole blood serum (WBS), defibrinogenated plasma, and plasma-derived serum (PDS). PDS obtained from platelet-poor plasma collected with the anticoagulant citrate phosphate dextrose and clotted by the addition of calcium chloride did not promote the growth of MCF-7 cells as well as did WBS or platelet-poor defibrinogenated plasma alone. Growth-inhibitory activity was observed when final cell densities in the presence of PDS combined with fetal bovine serum (FBS) were compared with those in control cultures maintained in FBS alone. The level of this activity in PDS varied among donors. Inhibition was not observed with HBL-100 cells. Calcium chloride did not induce inhibitory activity when added to WBS or defibrinogenated plasma, and platelet components did not alter the level of inhibition in PDS. However, platelet extracts did affect the expression of inhibition when added to plasma before clotting. Activity was diminished following dialysis of PDS, which suggested that inhibition stemmed from a small-molecular-weight factor for which expression depended on the mechanism of plasma coagulation.

Adenocarcinoma↗

Removal of terephthalic acid in alkalized wastewater by ferric chloride.

Terephthalic acid, which is a main component in alkali-decrement wastewater, is efficiently removed using ferric chloride in high pH solutions. About 90% removal of terephthalic acid is achieved at pH between 8 and 11. Especially, the removal reached 94.3% at pH 11. However, as the pH increased from pH 12 and 13, the low removal of terephthalic acid were found. The increasing ferric chloride dosage had a dramatic positive impact on the achieved removal of terephthalic acid. Further increase in the ferric chloride dosage did not produce better removal rate. The increase of terephthalic acid concentration also led to the increase of ferric chloride dosage in order to get the same removal of terephthalic acid. There was approximately a negative linear relationship between terephthalic acid concentration and removal of terephthalic acid. Compared with other coagulants, it can be seen that ferric chloride is more effective in a high pH solution and the amount of ferric chloride required is also less as compared with aluminum chloride, magnesium chloride and calcium chloride. Our results clearly showed that terephthalate anions strongly binds to positive Fe(OH)(3) flocs and forms insoluble complexes, probably through a mechanism involving electrostatic attraction. The electrostatic attraction may be particularly useful means of purifying wastewater in high pH solutions.

Adsorption↗

Noradrenaline activates a calcium-activated chloride conductance and increases the voltage-dependent calcium current in cultured single cells of rat portal vein.

1. Membrane responses were recorded by a patch pipette technique in cultured cells isolated from rat portal vein. Using the whole-cell mode, pressure ejections of noradrenaline evoked depolarization (current clamp) and inward current (voltage clamp) at membrane potentials of -60 to -70 mV. The noradrenaline-induced response was reversibly blocked by prazosin indicating that the response was mediated by alpha 1-adrenoceptors. 2. The ionic mechanism of the noradrenaline-induced inward current was investigated in potassium-free caesium-containing solutions. Alteration of the chloride equilibrium potential produced similar changes in the reversal potential of the noradrenaline-induced current, indicating that noradrenaline opened chloride-selective channels. There was no evidence implicating sodium or calcium as the charge-carrying ion. 3. Caffeine applied in the bathing solution also induced a transient increase in chloride conductance but the noradrenaline-induced response was lost after application of caffeine. This is interpreted to mean that the increase in chloride conductance induced by noradrenaline and caffeine can occur as a consequence of a rise in intracellular calcium concentration depending on release of calcium from the same intracellular stores. 4. In the presence of caffeine, noradrenaline increased both the voltage-dependent calcium and chloride membrane conductances during application of repetitive depolarizing pulses. It is concluded that in isolated cells of the rat portal vein the depolarization in response to noradrenaline is mediated by an increase in chloride conductance depending on both the calcium release from intracellular stores and the increase of the voltage-dependent calcium current.

Animals↗

Effect of mineral trioxide aggregate on proliferation of cultured human dental pulp cells.

AIM: To investigate the effect of mineral trioxide aggregate (MTA) on the proliferation of human dental pulp (HDP) cells ex-vivo. METHODOLOGY: Human dental pulp cells were cultured with MTA or calcium hydroxide-containing cement (Dycal) using culture plate inserts. Control cells were cultured with culture plate inserts only. Cell proliferation was measured for up to 14 days using a Cell Counting kit, and the concentration of calcium ions released from the tested materials was assessed using a Calcium E-test kit. To confirm that the effect of MTA was attributable to released calcium ions, cell proliferation was measured in the presence of exogenous calcium chloride as a source of calcium ions while in the absence of MTA. RESULTS: Mineral trioxide aggregate significantly stimulated cell proliferation after 12 days, whereas Dycal had no such effect. The number of calcium ions released from MTA was significantly higher than that released from Dycal. Following the addition of calcium chloride, cell proliferation increased in a dose-dependent manner after 12 days. Moreover, cell proliferation showed a similar pattern whether a given concentration of calcium ions was produced by calcium chloride or by release from MTA. CONCLUSIONS: In this ex-vivo study, the elution components such as calcium ions from MTA had higher proliferation ability of HDP cells than control and Dycal.

Adult↗

Inactivation of calcium-activated chloride conductance in Xenopus oocytes: roles of calcium and protein kinase C.

Inactivation of Ca2(+)-induced Cl- currents was studied in Xenopus oocytes using the two-electrode voltage-clamp technique. In oocytes permeabilized to Ca2+ by treatment with the ionophore A23187, Ca2+ influx caused by the addition of 2.5-5 mM Ca2+ to the extracellular solution elicited Cl- currents consisting of two components: a fast, transient one (Ifast) and a slow one (Islow). In response to a subsequent application of the same dose of Ca2+, Ifast and Islow were reduced (inactivation phenomenon). The inactivation did not depend on the direction of current flow, but did depend on the duration of the first exposure to Ca2+. The extent of inactivation of Ifast was more significant than that to Islow. Both Ifast and Islow fully recovered from inactivation in less than 30 min. Intracellular injections of 100-400 pmol CaCl2 evoked large inward currents but did not reduce the amplitude of currents evoked by Ca2+ influx. The activator of protein kinase C, beta-phorbol dibutyrate, caused full inhibition of Ifast without any change in Islow. H-7 (1,5-isoquinolinesulfonyl-1,2 methylpiperazine), an inhibitor of protein kinases, strongly reduced the extent of inactivation. Our results suggest that elevation of intracellular Ca2+ by Ca2+ influx through the plasma membrane causes inactivation of the Ca2(+)-dependent Cl- conductance via activation of a Ca2(+)-dependent protein kinase, possibly protein kinase C, whereas Ca2+ arriving at the membrane from inside the cell does not initiate the processes leading to inactivation.

Animals↗

Mechanism of the inhibitory action of indomethacin on smooth muscle.

1. Strips of muscle from the wall of the guinea-pig stomach contracted in response to electrical field stimulation (100 ms pulses, 0.2 Hz) or to histamine (1 mug/ml), and these responses were inhibited by indomethacin (2-20 mg/100 ml).2. Glycerol-extracted strips of stomach muscle developed tension when exposed to a mixture of ATP, CP, magnesium chloride and calcium chloride. The tension response was not altered by indomethacin (50 mg/100 ml).3. Indomethacin failed to alter the content of ATP or of CP in strips of stomach muscle.4. The calcium content of strips of stomach muscle increased in response to 30 min of electrical stimulation (100 ms pulses, 0.2 Hz). The uptake of calcium and the contraction of the strips were inhibited by indomethacin (2-20 mg/100 ml) to a similar extent.5. Calcium uptake by electrically stimulated guinea-pig aorta was inhibited by lower concentrations of indomethacin than were required by stomach muscle.

Adenosine Triphosphate↗

Analysis of calcium activated chloride current fluctuations in Xenopus laevis oocytes.

Fluctuations of calcium activated chloride currents were investigated in oocytes of Xenopus laevis. The method of noise analysis and the model of chloride channels activation by calcium ions were used to estimate the chloride channels lifetime and the average frequency of current fluctuations, which depends on changes of cytoplasmic calcium concentration. This current fluctuations can be evoked by activation of cholinergic receptors or inhibition by Na3VO4 of plasma membrane Ca(2+)-ATPase. The average opening lifetime of chloride channels was approximately 100 ms. The frequency of fluctuations increased with the increasing extracellular calcium concentrations and external ACh concentrations. Caffeine in 2 mmol/l concentration changed the current fluctuations into oscillations with a period of about 18-20s. Ten mmol/l caffeine fully inhibited the oscillation activity.

Action Potentials↗

Iontophoresis and chloride-containing compounds: parameters required for killing.

Fungi, and gram-positive and gram-negative organisms were susceptible to iontophoretic killing in simple media. Iontophoresis did not depend on electrode type but did require chloride-containing compounds in the medium. All organisms could be killed efficiently if chloride-containing compounds (for example sodium chloride and calcium chloride) were present in physiological concentrations. Effectiveness of iontophoretic killing could be reduced by nonphysiologically elevated concentrations of other substances (for example creatinine and albumin). The data suggest that iontophoresis should function well in urine, since chloride-containing compounds are present in adequate concentrations even if some naturally occurring compounds, such as creatinine or albumin, are elevated.

Calcium Chloride↗